Effect of pH and temperature on tropane alkaloids within a processing strategy to provide safe infant cereal-based food.

Tropane alkaloids (TAs) are secondary metabolites from weeds that can contaminate cereals and vegetables during harvest. Due to their toxicity, the Regulation (EC) 2023/915 sets maximum levels for atropine and scopolamine in cereal-based foods for infants containing millet, sorghum, buckwheat or their derived products. The aim of this study was to evaluate the effect of pH and temperature on the stability of TAs, as possible parameters in thermal processing to mitigate this chemical hazard in cereal-based infant food. The effect of pH (4 and 7) and temperature (80 °C and 100 °C) was assessed in buffer solutions. Also, treatment at 180 °C was performed in spiked and naturally incurred millet flour to assess the effect of high temperature, simulating cooking or drying, on the stability of TAs in the cereal matrix. The fate of 24 TAs was assessed by UHPLC-MS/MS. TAs showed high thermostability, although it was variable depending on the specific compound, pH, temperature and treatment time. In buffer solutions, higher degradation was found at 100 °C and pH 7. In spiked millet flour at 180 °C for 10 min, scopolamine and atropine contents decreased by 25 % and 22 %, similarly to other TAs which also showed a slow thermal degradation. Atropine, scopolamine, anisodamine, norscopolamine, scopine and scopoline were found in naturally contaminated millet flour. Interestingly, naturally incurred atropine was more thermostable than when spiked, showing a protective effect of the cereal matrix on TAs degradation. The present results highlight the need for an accurate monitorization of TAs in raw materials, as this chemical hazard may remain in infant cereal-based food even after intense thermal processing.

Determination of Atropine by HPLC in Plant of Datura by Liquid-Liquid Extraction and Magnet Solid-Phase Extraction.

Atropine is a tropane alkaloid found in abundance in Datura plant. We used two liquid-liquid extraction methods and magnet solid-phase extraction to compare the amount of atropine in these two types of plants (Datura innoxia and Datura stramonium). The surface magnetic nanoparticle Fe3O4 correction with an amine and dextrin, and finally, magnetic solid-phase extraction Fe3O4@SiO2-NH2-dextrin (MNPs-dextrin), was prepared. We determined the effect of significant parameters in the removal step and optimization of atropine measurements with half-fractional factorial design (25-1) and response surface methodology via central composite design. The optimum conditions are for desorption solvent = 0.5 mL methanol and desorption time of 5 min. We obtained an extraction recovery of 87.63% with a relative standard deviation of 4.73% via six frequented measurements on a 1 µg L-1 atropine standard solution based on the optimum condition. Preconcentration factors for MNPs are 81, limit of detection = 0.76 µg L-1 and limit of quantitation = 2.5 µg L-1.

Development, validation, and application of a multimatrix UHPLC-MS/MS method for quantification of Datura-type alkaloids in food.

A quantitative ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the determination of tropane alkaloids (TAs), atropine and scopolamine, in a variety of food products. The sample preparation of cereal-based food, oilseeds, honey, and pulses consisted of a solid-liquid extraction with an acidified mixture of methanol and water, while an additional step of solid-phase extraction on a cation-exchange sorbent was introduced in the treatment of teas and herbal infusions, aromatic herbs, spices and food supplements. The limits of quantification of the method varied from 0.5 to 2.5 µg kg-1. Apparent recovery was in the range of 70-120%, and repeatability and intermediate precision were below 20%. The method was successfully applied in a proficiency testing exercise as well as in the analysis of various commercial foods. Only 26% of the analysed food samples contained one or both TAs. The mean concentrations for atropine and scopolamine amounted to 21.9 and 6.5 µg kg-1, respectively, while the maximum concentrations were 523.3 and 131.4 µg kg-1, respectively. Overall, the highest levels of TA sum were found in an herbal infusion of fennel and a spice mix containing fennel and anise seeds.

Green extraction approach based on µSPEed® followed by HPLC-MS/MS for the determination of atropine and scopolamine in tea and herbal tea infusions.

A high throught methododology based on a green extraction technique, µSPEed®, followed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has been proposed for the analysis of atropine and scopolamine in tea and herbal tea infusions. For this, a digiVOL® Digital Syringe was used with different sorbents and working conditions to obtain a fast and efficient µSPEed® extraction. The best performance was achieved with a PS/DVB sorbent phase, sample loading of 5 × 500 µL and elution with 2 × 100 µL aliquots of methanol. The strategy based on µSPEed® followed by HPLC-MS/MS was validated, attaining quantitation limits lower than 0.15 ng mL-1 and recoveries between 94 and 106% for both analytes and applied to seventeen tea and herbal tea infusions. Fourteen infusions showed contamination with one or both analytes above the maximum content legislated (sum of atropine and scopolamine < 0.2 ng mL-1).

Scopolamine and atropine in feeds - determination with liquid chromatography mass spectrometry.

Tropane alkaloids (TAs) are naturally occurring plant toxins. Due to the fact that TA-producing plants can enter the food chain, they pose a risk for animals and human health. Therefore, sensitive analytical methods need to be developed to provide an adequate safety of feed and food. The presented method is based on liquid chromatography-mass spectrometry detection and enables the determination of scopolamine and atropine in compound feeds at a low level of contamination. Limits of quantification for scopolamine and atropine were 0.92 and 0.93 µg kg-1, respectively. Scopolamine-D3 and atropine-D3 were used for quantification. The method was successfully validated and applied to the analysis of 42 feed samples. Among investigated feeds, 67% contained at least one of the monitored alkaloids. Soybean meals were the feed materials contaminated most often, also with the highest determined concentrations of TAs, which reached 147.9 µg kg-1.

Development of CE-C4D Method for Determination Tropane Alkaloids.

A fast method for the determination of tropane alkaloids, using a portable CE instrument with a capacitively coupled contactless conductivity detector (CE-C4D) was developed and validated for determination of atropine and scopolamine in seeds from Solanaceae family plants. Separation was obtained within 5 min, using an optimized background electrolyte consisting of 0.5 M acetic acid with 0.25% (w/v) ß-CD. The limit of detection and quantification was 0.5 µg/mL and 1.5 µg/mL, respectively, for both atropine and scopolamine. The developed method was validated with the following parameters-precision (CV): 1.07-2.08%, accuracy of the assay (recovery, RE): 101.0-102.7% and matrix effect (ME): 92.99-94.23%. Moreover, the optimized CE-C4D method was applied to the analysis of plant extracts and pharmaceuticals, proving its applicability and accuracy.

Determination of atropine and scopolamine in spinach-based products contaminated with genus Datura by UHPLC-MS/MS.

Datura species are well known because of their high concentration of tropane alkaloids, which has led to poisoning episodes when Datura is accidentally mixed with edible crops. Therefore, the European Union has set a maximum level in cereal-based infant food products of 1 µg kg-1 for atropine and scopolamine. However, the occurrence of these compounds in other commodities has become a global concern. Spinach and derived products can be contaminated with Datura innoxia leaves. In this study, we tested frozen spinachs and spinach-based infant food products. The determination was carried out by UHPLC-MS/MS after applying the QuEChERS method as sample treatment. The LOQs were below 0.016 µg kg-1, achieving satisfactory results in terms of precision, accuracy, and matrix effects. The obtained results (ranging between 0.02 and 8.19 µg kg-1) were close to the maximum level set by the European Union for 24% of the samples tested.

An ultrasound assisted extraction-solid-phase extraction-ultra-performance liquid chromatography combined strategy for atropine determination in Atropa belladonna leaves.

Atropine is an antimuscarinic alkaloid identified in Atropa belladonna. In pharmacopeias, percolation is standardized as an extraction method for A. belladonna leaves, along with liquid-liquid extraction as a cleanup procedure and titration as an analytical method for assaying the atropine in the leaves. In this study, a faster, solvent-saving, and more reliable method for quality control of A. belladonna samples was developed. Ultrasound-assisted extraction was proposed and optimized by fractional factorial design followed by Box-Behnken design. For modeling atropine content, the following optimal conditions were established: particle size, 180 µm; percentage methanol in water, 50%; volume of solvent, 15 ml; time of extraction, 60 min; and number of extractions, two. This led to a significant improvement in atropine extraction (P < 0.001). For cleanup, solid-phase extraction was used as an alternative to liquid-liquid extraction, giving similar results, with higher reproducibility. Finally, for the atropine assay, a UPLC method was validated as a substitute for the classic titration method. Taken together, the development of an ultrasound-assisted extraction-solid-phase extraction-UPLC approach allowed the determination of atropine content in A. belladonna leaves in a time- and solvent-saving manner, with high reliability.

Indirect competitive enzyme-linked immunosorbent assay based on a broad-spectrum monoclonal antibody for tropane alkaloids detection in pig urine, pork and cereal flours.

In this study, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a broad-spectrum monoclonal antibody for tropane alkaloids (TAs) was established for the rapid screening of atropine, scopolamine, homatropine, apoatropine, anisodamine, anisodine and L-hyoscyamine residues in pig urine, pork and cereal flour samples through a simple sample preparation procedure. The half inhibitory concentrations of atropine, homatropine, L-hyoscyamine, apoatropine, scopolamine, anisodamine and anisodine were 0.05, 0.07, 0.14, 0.14, 0.24, 5.30 and 10.15 ng mL-1, respectivelyThe detection and quantitative limits of this method for TAs in samples were 0.18-73.18 and 0.44-74.77 µg kg-1. The spiked recoveries ranged from 69.88% to 147.93%, and the coefficient of variations were less than 14%. Good correlation (R2 = 0.9929) between the results of the ic-ELISA and the high performance liquid chromatography-tandem mass spectrometry support the reliability of the developed ic-ELISA method.

Application of a streamlined LC-MS/MS methodology for the determination of atropine and scopolamine in cereals from Asian and African countries.

Tropane alkaloids are toxic secondary metabolites produced by a wide variety of plants that can be present in edible materials or animal feed. Several human poisoning cases through consumption of cereals were reported over the last years and highlighted the need for reliable and robust analytical methodologies for safety control. To rationalize analyses in high-throughput laboratory environments dealing with shorter and shorter turn-around-around time, the scope of our multi mycotoxins method was extended to the analysis of two regulated tropane alkaloids, namely atropine and scopolamine. Extraction procedure is based on the QuEChERS (Quick, Easy, Cheap, Efficient, Rugged, and Safe) approach followed by liquid chromatography tandem mass spectrometry (LC-MS/MS) detection. Quantification is performed by the isotopic dilution approach using labelled isotopomers as internal standard. The procedure was validated at two fortification levels (0.5 µg/kg and 10 µg/kg) on different cereal-based products according to the European SANTE/12682/2019 document and performance parameters such as precision (RSD(r) ≤ 6%, RSD(iR) ≤ 6%) and recovery (82-114%) fulfilled its requirements. The limit of quantification (0.5 µg/kg) is low enough to ensure compliance with existing regulations. The method was further applied on 95 cereals and cereal-based products collected from Asian and African countries. All samples were found free of the two targeted TAs, with the exception of a rice-based product in which both atropine and scopolamine were quantified at 9.6 µg/kg and 2.6 µg/kg, respectively. A total of 29 cereals samples, shown to be free of both atropine and scopolamine were also analysed for mycotoxins. Aflatoxins, fumonisins, and deoxynivalenol were sporadically detected at levels below the maximum levels defined by the European Union legislation often considered as the most stringent regulation.

Determination of tropane alkaloids in cereals, tea and herbal infusions: Exploiting proficiency testing data as a basis to derive interlaboratory performance characteristics of an improved LC-MS/MS method.

Monitoring of tropane alkaloids is regulated in the European Union in cereal-based foods for infants and young children, tea and herbal infusions. The European Commission's Joint Research Centre (JRC) developed an improved LC-MS/MS analytical method using a pentafluorophenyl column, validated it and conducted two proficiency tests targeting these food categories. A subset of the data gathered from laboratories that used the JRC method was additionally exploited to derive interlaboratory performance characteristics. The method showed fit-for-purpose figures of merit. The LOQs for atropine and scopolamine were around 0.4 and 1.2 µg/kg in cereal products, and in tea and herbal infusions, respectively. Uncertainties varied from 15 to 25%. The reproducibility varied from 11 to 38% for scopolamine and from 17 to 44% for atropine at levels ranging from 0.18 to 18.8 and 1.2-54.0 µg/kg, respectively. Recoveries ranged from 71 to 96%. These performance parameters render the method a good candidate for standardisation.

Tale of Two Alkaloids: pH-Controlled Electrochemiluminescence for Differentiation of Structurally Similar Compounds.

Electrochemiluminescence (ECL) has increased in popularity as a result of its inherent advantages, including but not limited to portability, simplicity of use, and low reagent consumption. However, its significant advantages are often over shadowed as a result of its limited specificity. ECL emissions are intrinsically broad and lack the definition of other available analytical techniques. Furthermore, species with similar functional groups have almost identical electrochemical behavior and thus typically emit within approximately the same potential region. Within this contribution we have demonstrate the use of pH controlled ECL to prove the presence of two individual species within a mixed sample. Analysis at a single pH would not provide this information. We have illustrated the potential of this methodology to quantify scopolamine alongside sister tropane alkaloid atropine, a known ECL interferent. Previously the two alkaloids could not be distinguished from one another using a single technique which did not involve a separation strategy. pH controlled ECL is a simple approach to improve the specificity of a basic [Ru(bpy)3]2+ film based sensor. By exploiting molecular characteristics, such as pKa, we have been able to fine-tune our methodology to facilitate identification of analytes previously exhibiting indistinguishable ECL emission. Thus, by improving specificity, while maintaining operational simplicity and inexpensive design, we have been able to highlight the potential power of ECL for identification of structurally similar compounds. Further improvements of specificity, such as demonstrated within this contribution, will only further future applications of ECL sensors across a range of different fields.

Utilization of an Electrochemiluminescence Sensor for Atropine Determination in Complex Matrices.

A major challenge within forensic science is the development of accurate and robust methodologies that can be utilized on-site for detection at crime scenes and can be used for analyzing multiple sample types. The recent expansion of electrochemical sensors to tackle this hurdle requires sensors that can undergo analysis without any pretreatment. Given the vast array of samples that are submitted for forensic analysis, this can pose a major challenge for all electrochemical sensors, including electrochemiluminescent (ECL)-based sensors. Within this contribution, we demonstrate the capacity for an ECL-based sensor to address this challenge and it is potential to detect and quantify atropine from a wide range of samples directly from herbal material to spiked solutions. This portable platform demonstrates satisfactory analytical parameters with linearity across a concentration range of 0.75 to 100 µM, reproducibility of 3.0%, repeatability of 9.2%, and a detection limit of ∼0.75 µM. The sensor displays good selectivity toward alkaloid species and, in particular, the hallucinogenic tropane alkaloid functionality within complex matrices. This portable sensor provides rapid detection alongside low cost and operational simplicity, thus, providing a basis for the exploitation of ECL-based sensors within the forensic arena.

Toxicological analysis of cocaine adulterants in blood samples.

Cocaine was the second most widely used drug in Europe in 2016, with 3.5 million consumers aged 15-64 years old. Adulterants are pharmacologically active substances developed for medical purposes, however, there is little knowledge about their influence in the human body when there is concomitant use with cocaine. The objective of this work was to validate a method that allows the identification, confirmation and quantification of cocaine adulterants in blood samples collected in vivo or post-mortem. The studied substances were atropine, phenacetin, hydroxyzine, ketamine, lidocaine and tetramisole. A retrospective study of the prevalence of these substances, as well as their relative concentrations, was made analysing 97 real blood samples previously tested positive for cocaine and/or its metabolites. The analytes of interest were extracted, using a simple method based on protein precipitation with frozen acetonitrile and further analysis by GC/MS. The method was fully validated in accordance with parameters and criteria implemented in the lab and SWGTOX recommendations (mean recovery: 94-115%; CV: 6.2-13%; BIAS: 2.7-7.8%). 31 samples were positive for adulterants: phenacetin (19%), tetramisole (15%), lidocaine (8%) and hydroxyzine (1%). Concentrations were higher in post-mortem samples for all compounds analysed. Lidocaine was more prevalent in samples collected in vivo whereas tetramisole was present almost exclusively in post-mortem samples. Phenacetin was evenly distributed between post-mortem and in vivo samples. The validated method allows rapid, precise, accurate and economic analysis of selected compounds and requires smaller sample aliquots which can be important in post-mortem cases. The information collected can be important in future studies of correlation between the presence of adulterants and cocaine toxicity.

Reliable determination of tropane alkaloids in cereal based baby foods coupling on-line spe to mass spectrometry avoiding chromatographic step.

Cereal based foods are a major part of the infant diet and they can be contaminated with Solanaceae and other plants containing tropane alkaloids. This study was focused on the optimisation of an extraction procedure based on a solid-liquid method and an online SPE system, directly coupled to different mass spectrometry analysers as Orbitrap and triple quadrupole (QqQ), removing the chromatographic separation step. Total running analysis time was 15.8 min (17.3 min in the QqQ system). The developed method was validated obtaining recoveries ranging from 66 to 98% and 68-97% for the Orbitrap and QqQ respectively (RSD lower than 14.3%) and limits of quantification from 0.5 to 5 µg kg-1 and 0.5-10 µg kg-1 for the QqQ and Orbitrap analyser respectively. The validated method was applied to several cereal based baby food samples, finding a positive sample containing atropine (11.5 µg kg-1), scopolamine (2.8 µg kg-1) and apoatropine (7.5 µg kg-1).

MALDI imaging mass spectrometry revealed atropine distribution in the ocular tissues and its transit from anterior to posterior regions in the whole-eye of rabbit after topical administration.

It is essential to elucidate drug distribution in the ocular tissues and drug transit in the eye for ophthalmic pharmaceutical manufacturers. Atropine is a reversible muscarinic receptor used to treat various diseases. However, its distribution in ocular tissues is still incompletely understood. Matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS) evaluates drug distribution in biological samples. However, there have been few investigations of drug distribution in ocular tissues, including whole-eye segments. In the present study, we explored the spatial distribution of atropine in the whole-eye segment by MALDI-IMS, and then evaluated the changes in atropine level along the anterior-posterior and superior-inferior axes. A 1% atropine solution was administered to a rabbit and after 30 min, its eye was enucleated, sectioned, and analyzed by MALDI-IMS. Atropine accumulated primarily in the tear menisci but was found at substantially lower concentrations in the tissue surrounding the conjunctival sacs. Relative differences in atropine levels between the anterior and posterior regions provided insights into the post-instillation behavior of atropine. Atropine signal intensities differed among corneal layers and between the superior and inferior eyeball regions. Differences in signal intensity among tissues indicated that the drug migrated to the posterior regions via a periocular-scleral route. Line scan analysis elucidated atropine transit from the anterior to the posterior region. This information is useful for atropine delivery in the ocular tissues and indicates that MALDI-IMS is effective for revealing drug distribution in whole-eye sections.

Detection and identification of medically important alkaloids using the surface-enhanced Raman scattering spectroscopy.

Currently, trace detection of drugs, medicinal products, psychoactive substances, poisons and other natural or synthetic compounds in the human body has become one of the most important areas of interest in medicine, toxicology and forensic research. Due to the rapid development of nanotechnology, applications in forensic and biological sciences, food industry and art preservation there is an increasing interest in surface-enhanced Raman scattering (SERS) spectroscopy as a technique capable of low detection limits in the analysis of small amounts of studied analytes. In this study, different excitation wavelengths (785 nm and 1064 nm) were used to find the appropriate experimental conditions for the detection and identification of medically significant alkaloids - atropine and pergolide - by means of surface-enhanced Raman scattering spectroscopy. SERS spectra of selected alkaloids were measured in the concentration range 10-3-10-9 mol∙L-1 using large-scaled platinum substrates coated with electrochemically prepared gold or silver SERS-active layers. Identification was based on the assignment of surface-enhanced characteristic vibrational bands using theoretical (DFT) calculations and comparing them with normal (non-enhanced) Raman spectra of pure compounds. All sets of spectral data were subjected to multivariate statistical approach (partial least squares regression) aiming at prediction of alkaloids concentration in developed models and its comparison with experimental results.

An integrated approach to study novel properties of a MALDI matrix (4-maleicanhydridoproton sponge) for MS imaging analyses.

The chemical properties accounting for the operation of a valuable matrix used in matrix-assisted laser desorption ionization (MALDI) to perform mass spectrometry imaging (MSI), namely 3-(4,5-bis(dimethylamino)napthalen-1-yl)furan-2,5-dione (4-maleicanhydridoproton sponge, MAPS), have been elucidated also by comparison with the parent molecule 1,8-bis(dimethylamino) naphthalene (so-called proton sponge, PS). Both compounds present the bis(dimethylamino) groups, apt to efficiently trap a proton imparting positive charge. Only MAPS, though, owns the maleicanhydrido function acting as electrophile and yielding covalently bound adducts with a variety of analytes. In this way, MAPS performs as "carrier" for the analyte (A) of interest, at the same time minimizing the presence of useless, background ions. The covalent character of the adducts, [MAPS+H + A]+, is testified by their collision-induced dissociation pattern, quite distinct from the one displayed by [PS + H]+, while PS does not form any [PS + H + A]+, thus confirming the key role of the maleicanhydrido functionality of MAPS. Vibrational spectroscopy of [MAPS+H + A]+ adducts (A = H2O, NH3) provided further structural evidence. The presence of a mobile proton on A was found to be a requisite for adduct formation by electrospray ionization of acetonitrile solutions, pointing to a possible role of MAPS in discriminating competing analytes based on molecular features. The performance of MAPS has been verified in MALDI-MSI of Atropa belladonna berries, exploiting MAPS binding to atropine. Graphical abstract ᅟ.

Development and Validation of a High-Performance Thin-Layer Chromatographic (HPTLC) Method for Simultaneous Quantification of Reserpine, Atropine, and Piperine in Sarpagandha Ghanvati, a Classical Ayurvedic Preparation.

Background: Anxiety disorders are the most common of emotional disorders, affecting more than 20 million people annually. Sarpagandha Ghanvati is a classical Ayurvedic polyherbal formulation prescribed in conditions of insomnia, hysteria, and is used as an anxiolytic agent. Standardization and quality control are the two major issues that need to be addressed for herbal formulations, especially those containing multiple herbal ingredients. Objective: An HPTLC method was developed for the simultaneous quantification of reserpine, atropine, and piperine from Sarpagandha Ghanvati containing Rauwolfia serpentine (root), Hyoscyamus niger (seed), and Piper longum (root and stem). Methods: The marker compounds were effectively resolved on a silica gel G TLC plate using toluene-ethyl acetate-diethyl amine (7+2+1, v/v) as the mobile phase. The detected wavelengths for reserpine, atropine, and piperine were 269, 220, and 254 nm, respectively. The method was validated as per the International Conference on Harmonization guidelines. Results: R. serpentine roots contained 0.82% w/w of reserpine. Atropine content in the seeds of H. niger was found to be 0.004% w/w, whereas P. longum roots were found to contain 0.508% of piperine. The method was found to be accurate, which was evident from 98.93, 99.46, and 99.10% recovery of reserpine, atropine, and piperine, respectively, when the respective herbs were spiked with them. By the developed HPTLC method, 1.0 g of Sarpagandha Ghanvati was found to contain 4.94, 0.049, and 0.318 mg of reserpine, atropine, and piperine, respectively. The recoveries of these three markers from the formulation were found to be 90.32, 92.45, and 89.97%, respectively. Conclusions: The developed method can be successfully used for simultaneous estimation of these marker compounds and for the quality control of the classical Ayurvedic formulation Sarpagandha Ghanvati. Highlights: This works describes effects of extraction solvents on the quantities of marker compounds in the formulations. It also suggests a simple and reliable HPTLC method for simultaneous quantification of three different marker compounds from a poly-herbal formulation.

Simultaneous quantification of atropine and scopolamine in infusions of herbal tea and Solanaceae plant material by matrix-assisted laser desorption/ionization time-of-flight (tandem) mass spectrometry.

RATIONALE: Atropine (Atr) and scopolamine (Scp) are toxic secondary plant metabolites of species within the Solanaceae genus that can accidentally or intentionally reach the food store chain by inaccurate harvesting of any plant material, e.g. for herbal tea infusions. Ingestion may cause severe anticholinergic poisoning thus requiring risk-oriented determination in food and beverages. The suitability of matrix-assisted laser desorption/ionization time-of-flight (tandem) mass spectrometry, MALDI-TOF MS(/MS), should be characterized for simultaneous analysis. METHODS: We herein present the first MALDI-TOF MS(/MS) procedure for quantitative determination of both alkaloids in herbal tea infusions and Solanaceae plant material. A standard additions procedure using triply deuterated Atr as internal standard was developed and validated. RESULTS: Tropane alkaloids were detected without interferences and the standard additions procedure allowed reliable quantification. Intraday and interday precision were less than 17% and corresponding accuracies were between 77% and 112%. Limits of detection in the spotting solution were found at 5 ng/mL (Atr) and 0.5 ng/mL (Scp). The assay was applied to diverse tea infusions as well as to berries and leaves of deadly nightshade and angel's trumpet. CONCLUSIONS: The usefulness of MALDI-TOF MS(/MS) for investigations of plant-derived samples to prove contaminations by small basic compounds was demonstrated. The elaborate procedure is reliable but quite laborious to obtain quantitative results, but MALDI-TOF MS(/MS) was also shown to be a valuable tool for rapid qualitative screening for Atr and Scp in plant extracts.